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This diagnostic overlap necessitates a prompt and precise diagnostic tool for guiding accurate treatment and control. To address this diagnostic challenge, we developed basic recombinase polymerase amplification (basic-RPA) and RPA combined with lateral flow dipstick (RPA-LFD) assays for the detection of lactococcosis\n                      <jats:italic>-<\/jats:italic>\n                      associated\n                      <jats:italic>Lactococcus<\/jats:italic>\n                      species.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Both methods exhibited a detection limit of 10\n                      <jats:sup>3<\/jats:sup>\n                      copies\/\u00b5L for the recombinant plasmid pMD19-\n                      <jats:italic>adhE<\/jats:italic>\n                      -\n                      <jats:italic>ywdF<\/jats:italic>\n                      and 10 pg\/\u00b5L for\n                      <jats:italic>L. garvieae<\/jats:italic>\n                      genomic DNA. Specificity testing using DNA from\n                      <jats:italic>L. garvieae<\/jats:italic>\n                      ,\n                      <jats:italic>Streptococcus agalactiae<\/jats:italic>\n                      ,\n                      <jats:italic>Aeromonas hydrophila<\/jats:italic>\n                      ,\n                      <jats:italic>Edwardsiella piscicida<\/jats:italic>\n                      , and\n                      <jats:italic>Streptococcus iniae<\/jats:italic>\n                      confirmed that amplification occurred exclusively with\n                      <jats:italic>L. garvieae<\/jats:italic>\n                      , showing specificity against the tested panel. The assay also detected\n                      <jats:italic>L. petauri<\/jats:italic>\n                      , indicating that it recognizes both major lactococcosis-associated\n                      <jats:italic>Lactococcus<\/jats:italic>\n                      species. When tested with the plasmid standard, the RPA assays were 100-fold more sensitive than PCR; with genomic DNA, all three methods showed the same detection limit. When applied to 20 samples from experimentally infected tilapia, the RPA-LFD results were fully consistent with those of conventional PCR.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>The established methods are straightforward, sensitive, and specific, offering a promising candidate tool for the rapid on-site diagnosis of piscine lactococcosis.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12917-026-05587-5","type":"journal-article","created":{"date-parts":[[2026,5,28]],"date-time":"2026-05-28T10:53:29Z","timestamp":1779965609000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment of methods for visual and rapid detection of piscine lactococcosis based on isothermal recombinase polymerase amplification"],"prefix":"10.1186","volume":"22","author":[{"given":"Hanyu","family":"Luo","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhihao","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weiqi","family":"Fan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Zeng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong-An","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,5,28]]},"reference":[{"issue":"2","key":"5587_CR1","doi-asserted-by":"publisher","first-page":"436","DOI":"10.3390\/microorganisms13020436","volume":"13","author":"I Sirakov","year":"2025","unstructured":"Sirakov I, Strateva TV, Boyanov VS, Orozova P, Yordanov D, Rusenova N, et al. 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